Figure 1.
Characterization of hMSCs by flow cytometric analysis.
(A) Isolated hMSCs were positive for surface markers CD146, CD29, CD147 and CD44, and negative for CD45 and CD34. (B) The individual percentages of each surface marker expressed in these cells from the quantitative FACS analysis. Data represent the mean ± SD (n = 3).
Figure 2.
Sulfated GAG (sGAG) content normalized to wet weight in 3D pellet culture on Day 10, 17 and 24, with and without HS (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Data represent the mean ± SD (n = 4). P values were calculated using student t-test.
Figure 3.
Representative images of immunohistochemical staining of collagen type II in pellet culture samples on (A) Day 17 (B) Day 24.
Scale bar: 50 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Figure 4.
Representative images of immunohistochemical staining of aggrecan in pellet culture samples on (A) Day 17 (B) Day 24.
Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Figure 5.
Representative images of immunohistochemical staining of collagen type I in pellet culture samples on (A) Day 17 (B) Day 24.
Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Figure 6.
Representative images of immunohistochemical staining of collagen type X in pellet culture samples on (A) Day 17 (B) Day 24.
Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Figure 7.
Representative images of immunohistochemical staining of inducible heat shock protein 70 (HSP70) in pellet culture samples on (A) Day 17 (B) Day 24.
Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).
Table 1.
Semi-quantified Immunohistochemical staining Intensity.
Figure 8.
Western Blot analysis of collagen type II, aggrecan, and HSP70 expression in 3D chondrogenic pellet cultures using hMSCs from the 24 year old donor at Day 17 and Day 24.
(A), (C), (E), (G), (I) and (K) are the images of Western blot membranes while (B), (D), (F), (H), (J) and (L) are their semi-quantified band intensities respectively (Chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).